A fluorometric method for determining the degree of biotinylation of proteins.
A method to determine the number of biotin moieties attached to a protein has been developed based on quenching the natural fluorescence of avidin or streptavidin by biotin. The assay consists of titrating the number of biotin combining sites on streptavidin/avidin before and after adding the biotinylated protein. With this method only those biotin moieties capable of binding to streptavidin/avidin are detected. The assay is simple and sensitive, requiring only 1-10 micrograms of biotinylated protein per determination.
Der-Balin
G P
GP
Molecular Devices Corporation, Menlo Park, CA 94025.
Gomez
B
B
Masino
R S
RS
Parce
J W
JW
D000818:Animals; D001360:Avidin / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001710:Biotin / Q000378:metabolism*; D015200:Fluoroimmunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D019809:Streptavidin
D001426:Bacterial Proteins; D007074:Immunoglobulin G; D011506:Proteins; D001360:Avidin; D001710:Biotin; D019809:Streptavidin